摘要目的 检测糖尿病(DM)和糖尿病肾病(DN)患者血清lncRNA-生长停滞特异基因5 (lncRNA GAS5)和微小RNA-21 (miR-21)的表达变化,分析其与临床和病理指标的相关性,明确lncRNA GAS5和miR-21对DM和DN的诊断效能.方法 入组人群分为3组:DN组(肾活检病理诊断明确,n=25,男14例,女11例)、DM组(尿微量白蛋白/尿肌酐正常,n=10,男4例,女6例)和正常对照组(NC组,n=9,男4例,女5例).空腹采血,分离血清,实时荧光定量PCR法检测基因表达.应用t检验、Pearson、Spearman检验及多元线性回归分析lncRNA GAS5及miR-21表达水平和各临床指标的相关性,单因素方差分析检验lncRNA GAS5及miR-21在不同分组中的差异性,用受试者工作特征(ROC)曲线下面积(AUC)评价lncRNA GAS5及miR-21在疾病中的诊断效能.所有数据采用SPSS 20.0统计,GraphPad软件处理,P＜0.05视为差异有统计学意义.结果 (1)DM和DN患者血清lncRNA GAS5的表达水平均低于正常对照组,血清miR-21的表达水平均高于正常对照组,差异有统计学意义(均P＜0.05).(2)DN患者中,随着24 h尿蛋白量的逐渐增多,血清lncRNA GAS5的表达逐渐上调.随着肾活检病理分级的进展(Ⅱb-Ⅲ期),血清miR-21的表达也上调(P＜0.05).(3)空腹血糖(FBG)和糖化血红白蛋白(HbA1c)与血清lncRNA GAS5呈负相关(P＜0.05),并且FBG与血清lncRNA GAS5独立相关(P＜0.05);尿微量白蛋白(MA)、总胆固醇(TC)、Ser、BUN和SBP与血清miR-21水平呈正相关(P＜0.05);血清白蛋白(ALB)和肾小球滤过率(eGFR)与血清miR-21水平呈负相关(P＜0.05),并且ALB与血清miR-21独立相关(P＜0.05).(4)血清lncRNA GAS5、miR-21及lncRNA GAS5/miR-21“诊断标签”诊断DM的价值较好(P＜0.05).(5)血清miR-21及lncRNAGAS5/miR-21“诊断标签”诊断DN的价值较好(P＜0.05).结论 (1)血清lncRNA GAS5在DM中有较好的诊断效能.lncRNA GAS5/miR-21“诊断标签”诊断DM灵敏度85.71％,特异度88.89％,可以作为诊断DM的生物标志物.(2)血清miR-21表达水平可以作为DM和DN肾病的无创性诊断标志物.

abstractsObjective To elucidate the efficiency lncRNA GAS5 and miR-21 as biomarkers in diabetes mellitus and diabetic nephropathy.Methods The patients were divided into three groups,diabetic nephropathy group (DN group proven by renal biopsy,n=25,14 males and 11 females),diabetes group (DM group,with normal urine albumin creatinine ratio,n=10,4 males and 6 females),and normal control group (NC group,n=9,4 males and 5 females).The expressions of lncRNA GAS5 and miR-21 in serum samples were detected by real-time quantitative PCR.The correlation between serum lncRNA GAS5 and miR-21 expressions and the clinical parameters was analyzed by T-test,Pearson,Spearman test and multivariate linear regression analysis.Differences of lncRNA GAS5 and miR-21 in different groups were analyzed by one-way analysis of variance.The ROC curve was used to analyze the diagnostic efficacy of lncRNA GAS5 and miR-21 in diabetes and diabetic nephropathy.All data were analyzed by SPSS 20.0 and GraphPad software,with P ＜ 0.05 as considered statistically significant.Results (1) The expression of serum lncRNA GAS5 was significantly down-regulated and serum miR-21 was significantly up-regulated in both diabetes mellitus and diabetic nephropathy patients compared to the NC group all (P ＜ 0.05).(2) In DN patients,the expression of serum lncRNA GAS5 was gradually up-regulated along with the increment of 24 h urinary protein.The expression of serum miR-21 was gradually up-regulated along with renal biopsy stage Ⅱb-Ⅲ of DN (P ＜ 0.05).(3)FBG and HbA1c were all negatively correlated with serum lncRNA GAS5 (P ＜ 0.05),and FBG was independently correlated with serum lncRNA GAS5 (P ＜ 0.05).Urine microalbumin,Total cholesterol (TC),Scr,Urea and SBP were all positively correlated with serum miR-21(P ＜ 0.05).Albumin (ALB)and estimated GFR (eGFR) were negatively correlated with serum miR-21(P ＜ 0.05),and ALB was independently correlated with serum miR-21 (P ＜ 0.05).(4) The diagnostic efficiency of serum lncRNA GAS5,miR-21 and lncRNA GAS5/miR-21 as "diagnostic signature" for DM were was good (P ＜ 0.05).(5) The diagnostic efficiency of serum miR-21 and lncRNA GAS5/miR-21 as "diagnostic signature" for DN were was good (P ＜ 0.05).Conclusions (1) Serum lncRNA GAS5 had good diagnostic efficiency in diabetes mellitus.The sensitivity of lncRNA GAS5/miR-21 for diagnosis of diabetes was 85.71％,and specificity was 88.89％.(2) The level of serum miR-21 can be used as a noninvasive diagnostic marker for diabetic nephropathy.